| Project/Area Number |
15590889
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurology
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| Research Institution | National University Corporation Tottori University |
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Principal Investigator |
ARAGA Shigeru Tottori University, faculty of Medicine, Associate Professor, 医学部, 助教授 (40193065)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | KM mouse / complementary peptide / myasthenia gravis / idiotype antibodies / anti-idiotype antibodies |
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| Research Abstract |
KM mice were immunized with the complementary peptide (RhCA 67-16 peptide) for AChR61-76 peptide which was B cell epitope of myasthenia gravis(MG). KM mouse can produce human immunoglobulins, not mouse immunoglobulins. We obtained four different subclasses of IgG monoclonal antibodies which can recognize the idiotype in myasthenia gravis(MG). Using these monoclonal antibodies, we have developed new diagnostic method for detecting the idiotype antibodies in MG patient sera.
Biotin labeled complementary peptide antibodies were fixed with streptavidin coated 96-well ELISA plates. Then, serial diluted MG serum samples were incubated on the plate. Captured idiotype antibodies were eluated with acidic buffer. IgG concentrations in the eluated solution were measured in sandwich ELISA system.
Forty two MG samples (male 13 examples, woman 29 examples) and 37 normal control samples (male 15 examples, woman 22 examples) were analyzed using these systems. The normal upper limits were determined using mean+3SD in the normal controls. The samples above the upper limits were counted as a positive one. Fourteen out of 42 MG samples were positive. All positive samples were seropositive MG samples. However, none of the seronegative MG samples were detected using these systems.
Complementary peptide antibodies using KM mouse will provide not only the new diagnostic tools but also the new therapeutic approach for MG patient.
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